(Iris Aurantica) in Syria

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Wild Golden Iris (Iris aurantica) in Syria

Khalil AL Maarri*, Thuraya Abou zedan and Nabil AL Batal Faculty of Agriculture, Damascus University, Syria Submission: February 14, 2017; Published: February 27, 2017 *Corresponding author: Khalil AL Maarri, Damascus University, Faculty of Agriculture, Damascus, Syria, Email:

Abstract

Iris aurantica

is a rhizomatous perennial, from the south mountain of Syria (JabalAl Drouze), it has a compact rhizomein vitro and propagation falcate leaves. and conservation,It has a slender and stem cryopreservation with flowers golden of Iris yellowaurantica to coppery-brown. The golden iris in Syria is becoming rare due to destruction of their natural habit. In this paper, plant classification, botanical description, morphological, geographical, chemical composition, Keywords: are investigated.

Abbreviations:Morphological; Botanical description; Micro propagation; Conservation; Chemical composition

MS: Murashige and Skoog; HF-MS: Hormon Free Murashige and Skoog; PVS2: Plant Vitrification Solution 2; LN: Liquid Nitrogen

Introduction urbanization, migration, determental climatic and environmental changes, adding the huge destruction of plant biodiversity by the endemic species, some of which belong to genus Lilium, Crocus, Syrian flora has 3247 species [1]. There are many of

hard war since 5 years [6]. Tulips and Iris. Iris is the largest and most complicated genus morphological, geographical, chemical composition, genetic of iridaceae, which includes over 300 species [2]. This includes In this paper, Plant classification, botanical description, variability, in vitro propagation and conservation, and some of the world most popular and varied garden flowers that cryopreservation of Iris aurantica are originatedIris in bothis considered Japan and as the a Mediterraneanwild perennial [3].herbaceous were investigated. In Syria, Scientific Classification plant that subjected to strict protection, though Iris grows Iris aurantica L is one of the important species that belongs naturally in many regions of Syria. It presents some 30 species to the family Iridaceae, endemic to Jabal Al `Arab, rhizomatous Apogon, Pogonias, Xiphion, Guno and oncocyclus which includes grown in Syria [1]. There are five subgenus found in the world. most of the Syrian species, that are considered as rare endemic plants, characterized by special, beautiful forms that have a great (with thick, creeping underground stems) [7]. importance in applied studies for genetic biodiversity, such as A. Kingdom: Plantae. Asparagales Iris aurantiaca B. Unranked: Angiosperm, Monocots. Order irideae. Iris aurantica Dinsm. C. Family: Iridaceae. Subfamily: Iridoideae, Tribe: Iris. , of Syria was first discovered by Dinsmor aurantica. on the Tell Quleib in Syria. Mouterde found it in several other D. Genus: Iris, Subgenus: Oncocyclus. Species: places in Djebel Druze, Tell Qouleib, Kafer, Tell Jaffna, Mayamas, Description of Iris aurantica Sahwet-El-Khodr [1] and Distribution in the Djebel Druze at Iris aurantica is a perennial plant, growing from compact about 1600m [4]. Dr. Werkmeister, Professor of Botany at the Botanical Institute, Geisenheim am Rhein, Germany which had the opportunity to collecting the golden Iris in 1961,and cultured rhizome that reaches up to 10-14 cm long. Rhizome develops it in his garden. The golden Iris flowered in Europe in June one from axillary buds, it allows new shoots to grow upwards. The golden iris rhizomes planted underground about 5-10cm in month later than in their natural habit in Syria [5]. proteins, starches and lipids, these nutrients become useful for destruction of their natural habit, as well as over harvesting rocky soils (Figure 1). It uses rhizomes to store nutrients like Iris plants in Syria are becoming rare due to both ongoing

the plant when new shoots will be formed in early spring. of wild species and the influencing of modernization, i.e., Curr Trends Biomedical Eng & Biosci 1(5): CTBEB.MS.ID.555573 (2017) 00111 Current Trends in Biomedical Engineering & Biosciences

golden yellow to coppery-brown, 8-9cm long, 5-5.5 wide, with 4cm wide, abovate, with minute purplish red spots and very fine purple veins. The falls are oblong-shaped, 7cm long, and

fine reddish veins. It has golden yellow with fine purple. Style branches (stigma) keeled, and have lobes tips that are a similar color like falls (Figure 3). After the iris has flowered, it produces a seed capsule, 8cm long, rather narrow.

The golden iris flowered in Europe in June, a whole month later than in their habitat in Syria. The unexpectedly cold European weather in the spring of 1962 had nearly everywhere Figure 1: Rhizome of iris aurantica. in Europe killed the flowers on the iris species. The only exception and a unique consolation, was the plants of Iris aurantica from GoldenSyria, which Iris gave Propagation us much joy with their flowers [5]. Iris species is usually accomplished vegetatively through bulbs or splitting of rhizomes (rhizomatous The propagation of Iris Iris, splitting the rhizomes gives a

). In rhizomatous the propagation of Iris maximum of 10 plants per year per rhizome [8]. Furthermore, species through seedlings is known to be difficult due to a poor fruit set and a very low germination rate. propagation and conservation of plants, especially for those that Plant tissue culture is a powerful alternative technique for

are rare and difficult to propagate by conventional methods. Figure 2: Leaves and roots of iris aurantica. Therefore, a new trend has evolved to propagate these species deterioration and to study the possibility of using them as a through tissue culture techniquein order to preserve it from Iris aurantica has green falcate leaves, they can grow up to medicalMicro or propagation ornamental plant.is the aseptic culture of cells, pieces of

15-25cm long and between 1-1.5cm wide (Figure 2). Leaves can small pieces of plant tissue identical to the plant from which it sheath up to haft of the stem after the iris has bloomed. At the tissue, or organs. It is possible to regenerate new plants from summer sun with dry conditions, leaves fade and die. Golden Iris has a slender stem, which can grow up to 30-50cm tall. The was derived. stages: stems hold terminal flowers (top of stem), blooming during May The process of micro propagation can be divided into four and Jun, they can flower for up of month long [4]. A. Initiation stage:

The objective of this stage is to achieve an aseptic culture. An aseptic culture is one without of rhizomes in Iris aurantica (after surface disinfection by contaminating bacteria or fungi. Base of leaves and shoot tips

chlorox 3%) were cultured on solidified MS medium containing 30g/l sucrose, and supplemented with 2mg/lBAP and 0.2mg/ month of culture, that using shoot tips of rhizomes resulted in lIBA (Abouzedan and Al-Batal 2015). Results showed, after one

the highestB. Multiplication growth percentage stage: (35.76%) A growing in initial ex plantstage [9].can be Figure 3: Iris aurantica flower parts. Iris aurantica, the highest induced to produce vegetative shoots by including a cytokinin Golden Iris has a slender stem, which can grow up to 30- in the medium and different media. In average number of shoots per ex plant was found (3.43) under 50cm tall. The stems hold terminal flowers (top of stem), resulted in the highest multiplication rate and shoot length with blooming during May and Jun, they can flower for up of month BAP at a concentration of 3.0mg/l (Figure 4), and MS media long [4]. The flowers like other irises, it has two pairs of petals, of the plantlets on the same medium resulted in increasing significant difference compared with Heller media, Subculture three large sepals, known as the falls and three inner smaller multiplication rate and shoot length in Iris aurantica petals or sepals, known as the standards. The standards are [9] the How to cite this article: Khalil AL M, Thuraya A z, Nabil AL B. Wild Golden Iris (Iris aurantica) in Syria. Curr Trends Biomedical Eng & Biosci. 2017; 1(5): 00112 555573. Current Trends in Biomedical Engineering & Biosciences

acclimatization in vivo was achieved easily with high percentage treatment of high concentrations of cytokinins (5 and 10mg/ tissue-cultured plants are extremely susceptible to wilting. The iris aurantica lBAP) consist of appearance of vitrification. later, plantlets were cultured in greenhouse and the average of success (86.95%) in (Figure 6). Two months,

length of shoots were 23. 25 cm [10].

Figure 4: Multiplication stage in Iris aurantica. Figure 6: Acclimatization stage. A: plantlet after one week, B: plantlet after 2 months, C: plantlets Growing shoots can be induced C. Rooting stage: in greenhouse. to produce adventitious roots by including an auxin in the In iris aurantica In Vitro Conservation medium. , the highest root percentage (88.5%) was obtained on medium containing 3mg/lIBA (Figure 5). conservation plants material for periods ranging from 6 months Conservation is a very simple in vitro technique that permits The highest root number (4.25) was recorded when using the concentration 0.5mg/lIBA [9] (Table 1). on reducing the growth rates of the tissue cultured plant and to 5-7 years, depending on species [11]. This technique is based was conducted, to develop an in vitro yet increasing the intervals between subcultures [12]. Research conservation and relieve of growth and increase the period technique for short-term of time between transfers of Iris aurantica agents for in vitro conservation was sucrose compared with , The best osmotic iris aurantica (mannitol and sorbitol) and the best medium concentration was 1/10 MS [10] in the cultured stored at (3 ˚C) gave the highest survival (93.33%) and lengthening the time to regulate expression of many genes that are responsible period between transfers up to 6 months [10]. ABA was found for the syntheses of proteins needed for osmotic adjustment -3 Figure 5: Rooting stage, 1mgdm IBA. in the cell, such as, membrane stabilization proteins, and the LEA proteins that would modify water state in the cell to cope Table 1: The best media of tissue culture in iris aurantica. Compositions of Initiation Multiplication Rooting is responsible for low temperature tolerance capacity of plant with osmotic stress [13]. Some researchers reported that ABA Growth Medium Stage Stage Stage aurantica micro shoots were Medium MS MS ½MS conserved for more than nine months at normal growth room Sucrose tissues [14]. In our research Iris -3 -3 -3 Agar 30g7g dm dm 30g7g dm dm 30g7g dm dm -3 -3 -3 conditions in the media supplemented with 0.5 to 3.0mg/lABA, 1mg dm 1mg dm 1mg dm also ABA significantly decreases growth of shoots in medium -3 -3 -3 Thiamine(vitamin when compared with control. B1) 100mg dm 100mg dm 100mg dm nine month were obtained in media supplemented with 2 and -3 -3 -3l Results showed that the best survival after three, six and Myo- inositol (B8) Hormones 2mg dm 0 -3 -3 3mg/l ABA with significant difference compared to the control BAP 3.0mg dm -3 -3 -3 at 3 ˚C and normal growth room conditions. The best treatments Other Compounds IBA 0.2mg dm 0.1mg dm 3.0mg dm on germ plasm conservation were in 3mg/l ABA at 3 ˚C, in these Activated Carbon - - treatments the survival rate was 60%. -3 Long - Term Conservation (Cryopreservation) D. Acclimatization:3g dm A growing, rooted shoot can be done, the humidity must be gradually reduced over time because The principle of cryopreservation is the storage of plant removed from tissue culture and placed in soil. When this is material at ultra low temperature (-196 ˚C) that takes a place in a cryogenic condition which is liquid nitrogen [15]. At this

How to cite this article: Khalil AL M, Thuraya A z, Nabil AL B. Wild Golden Iris (Iris aurantica) in Syria. Curr Trends Biomedical Eng & Biosci. 2017; 1(5): 00113 555573. Current Trends in Biomedical Engineering & Biosciences

temperature, all forms of cellular divisions and metabolic were demonstrated to have pesticide, anti neo plastic and anti medical use. The isolated compounds from some species activities of plant cell are ceased and consequently plant material can be stored unaltered for an indefinite time scale [16,17]. Two tuberculosis properties [27]. The extract of I. germanica was iris: techniques of cryopreservation were used in the case of Golden found to have central anti serotonin activity [28]. Vitrification Gas chromatography–mass spectrometry (GC-MS) analyses in Iris germanica Iris aurantica of the essential oil have indicated the presence of 23 compounds , and 19 in [29]. The major Vitrification is based on three major phases, the loading Iris germanica, and Iris aurantica respectively with no phase, dehydration with the highly concentrated vitrification compound in these essential oils was Myristic acid (61.42%, solutions, and unloading phase [18]. In loading phase samples 70.67%) in significant differences [29]. The findings here agreed with those are exposed to cryo protectants or diluted vitrification solutions compound of the oil of the fresh and naturally aged rhizomes obtained [30] which noted that the myristic acid was the major [19] the samples are dehydrated by a highly concentrated in Iris pallida vitrification solution before being plugged in LN, [20]. In cryovials after rapid thawing, and then replaced routinely with sub major compounds obtained were Lauric acid, Decanoic unloading phase vitrification solution is drained out of the which has antifungal properties [31]. The other

1.2 M sucrose for 10-20 min [21]. Plant vitrification solution 2 acid (Capric acid), Palmitic acid methyl ester, Octadecanoic acid (PVS2) is commonly used in most vitrificationIris aurantica protocols were [22].excised methyl ester, Elaidic acid methyl ester (9- Octadecenoic acid methyl ester (E) and Palmitic acid [29]. In our research Shoot-tips in in I. aurantica The highest percentage of Lauric acid was obtained (6.97%) aseptically from in vitro grown plants and incubated for 3 days Iris germanica , with no significant differences comparing with on solid (HF-MS) media supplemented with 0.3 M sucrose under investigate the use of iris aurantica complete darkness at 24 ± 1˚C.in vitrification, shoot-tips were (5.69%), these findings give us the possibility to desiccation with different combinations and concentrations of loaded in 0.4 M sucrose and 2 M glycerol for 20 min followed by Iris Cultivation for medical purposes [29]. I. Iris aurantica aurantica PVS2, before immersion in LN. Results showed that Subjecting Wild is very difficult to cultivate. It can shoot-tips to gradual increase in PVS2 concentrations withstand the cold and the heat as long as it is dry. Golden iris (20, 40, 60, and 100%) before plunging them in LN gave the needs well drained soil and at least 6-8 hours sunlight. If the soils highest survival for the non-cryo preserved shoot-tips (50%), are heavy, sand or humus may be added to improve drainage. and cryo preserved shoot-tips ( 30%), respectively, Using of 2 Iris The ideal pH is less than 7, slightly acidic. Iris should be planted M glycerol plus 0.4 M sucrose resulted in the highest survival aurantia in September or October, when the weather starts to cool, it is (80%, 25%) for the non cryo preserved and cryo preserved non cryo preserved I. aurantica preferably to be divided and planted at least six weeks before the shoot-tips, respectively, The highest survival for the first frost in any area. shoot-tips was recorded after 30 minEncapsulation-Dehydration of loading. The rhizomes produce more rhizomes, which in turn lead to more leaves and flowers. One rhizome of golden iris can give it is necessity to divide the plants, removing and replanting the embryos or callus cells are encapsulated within alginate beads more than ten flowers. When the bloom production slows and This method, developed [23] in which shoot tips, somatic baby rhizomes in spacing 15-25 cm. Close planting results in and subsequent culture in a medium containing elevated immediate effect, faster clump formation, and makes dividing beads are then allowed to dehydrate using silica gel or by air irises need moisture and fertilizer to help their root systems concentrations (0.7-1.5M) of sucrose for 1 to 3 days [24,25]. The clumps a necessity in two to three years. New plantations of under the laminar air flow until the moisture content drops to becomes established. Watering depends on the soil quality and were encapsulated 20-30% before being immersedIris in auranticaLN [23]. the climatic conditions (Figure 7). It is preferable to give deep In our study shoot-tips in watering at long interval is better than shallow watering. in 3% calcium alginate and dehydrated under laminar air flow cabinet for0, 2, 4, or 6h. the highest survival were obtained after pre-treating encapsulated non-cryo preserved shoot-tips for 3 Chemicaldays in 0.5M Analysis sucrose supplemented media with 4h dehydration.

The medicinal parts of Iris (orris) species are the rhizomes with the roots. They contain volatile oil (α, β, γ, irons) giving the Figure 7: Soil of Mayamas Al Suwayda Syria. odor of violets triterpenes, isoflavonoids, flavonoids, xanthones and starch [26]. Some of Iris species were reported for their

How to cite this article: Khalil AL M, Thuraya A z, Nabil AL B. Wild Golden Iris (Iris aurantica) in Syria. Curr Trends Biomedical Eng & Biosci. 2017; 1(5): 00114 555573. Current Trends in Biomedical Engineering & Biosciences

References 17. 1. Shatnawi MA, Anfoka G, Shibli RA, Al-Mazra’awi MS, Shahrour W, et al. (2011) Clonal Propagation and cryogenic storage of virus free Moutrde P (1966) Nouvelle flore du Liban et de la Syrie. Del, Impri grapevines (Vitis vinefera L.) via meristem culture. Turk J Agric For 2. Catholique Beyrouth, Liban. 18. 35: 173-184. Trease WC (2002) Trease and Evan Pharmacognosy. W B Saunders, Ashmore SE (1997) Status report on the development and application 3. Edinburgh, London, New York, USA, pp. 26. of in vitro techniques for the conservation and use of plant genetic 4. Rasoul TN (1984) Ornamental bulbs. Al-Mousel University, Iraq. 19. resources. Rome. Dykes WR (1997) A Guide to Species Irises: Their Identification and 20. Withers LA (1991) In vitro conservation. Biol J linn Soc 43: 31-42. Cultivation, edited by the Species Group of the British Iris society. Sakai A, Kobayashi S, Oiyama I (1991) Survival by vitrification of 5. °C Cambridge University, pp. 424. nucellar cells of navel orange (Citrus sinensisvar. brasiliensis Tanaka) Werckmeister P (1963) The Iris aurantica of Syria. Bulletin of the 21. cooled to -196 . J Plant Physiol 137: 465-470. 6. American Iris society. 169: 29-30. genetic variability, propagation, conservation, active compound and Shibli R A, Shatnawi M, Subaih W, Ajlouni M (2006) In vitro conservation Al Maarri K (2016) Final report of project wild Iris in Syria: study of and cryopreservation of plant genetic resources: A review. World J 22. Agric Sci 2: 372-382. 7. biological effect. High commission scientific research, pp. 115. Reed BM (2008) Plant cryopreservation: A practical guide. USDA-ARS Mathew B (1989) The Iris. Oregon Timber Press, Portland, Oreg, pp. 23. National Clonal Germplasm Repository. Corvallis, USA, 3(11): 33-41. 8. 326. Fabre J, Dereuddre J (1990) Encapsulation-dehydration: a new Jehan H, Courtois D, Ehret C, Lerch K, Petiar V (1994) Plant regeneration approach to cryopreservation of Solanum shoot tips. Cryo Letters 11: of Iris pallid Lam and Iris germanica L via somatic embryogenesis from 24. 413-426. 9. leaves, apices and young flowers. Plant Cell Rep 13(12): 671- 675. Gonzalez-Arnao MT, Engelmann F (2006) Cryopreservation of plant Abouzedan T, AlBatal N, Al Maarri K (2012) In vitro micro propagation germ plasm using the encapsulation-dehydration technique: Review and chemical analysis of some wild Iris species in Syria. Master of 25. and case study on sugarcane. Cryo Letters 27(3):155-168. 10. Cryopreservation of apices of in vitro plantlets of almond (Prunusdulcis Faculty of Agriculture, Damascus University, 99 pages. Shatnawi MA, Engelmann F, Frattarelli A, Damiano C (1999) Abouzedan T, AlBatal N (2016) In vitro Micro propagation and Conservation of Iris aurantica L in Syria. The Arab Center for the 26. Mill.). Cryo Letters 20(1): 13-20. 11. Studies of Arid Zones and Dry lands (ACSAD). Guvence A, Kurucu S, Koyuncu M, Arihan O, Erdurak CS (2005) El-Dawayati MM, Zaid ZE, Elsharabasy SF (2012) Effect of Conservation Investigation on the seeds of Iris spuria L Subsp musulmanica (fomin) on Steroids Contents of Callus Explants of Date Palm cv. Sakkoti Aust J 27. takht (Iridaceae). Turkish J Pharm Sci 2(3): 125-136. 12. Basi Appl Sci 6(5): 305-310. Deng G, Zhang H, Xue H, Chen SH, Chen X (2009) Chemical composition Engelmann F (1991) In vitro conservation of tropical plant germplasm and biological activities of essential oil from the rhizomes of Iris 13. - a review. Euphytica 57(3): 227-243. 28. bulleyana. Agric Sci China 8(6): 691-696. Taiz L, Zeiger E (2002) Plant Physiology. 3rd edn, Sinauer Associates Al Maarri K, Abouzedan T, AlBatal N (2013) Chemical analysis of 14. Pubilsher, Hardcover. essential oils of some syrian wild Iris species. Am J Biochem Mol Biol 29. 3(1): 38-49. Popova EV, Lee EJ, Wu CH, Hahn EJ ,Paek KY (2009) A simple method for cryopreservation of Ginkgo bilobacallus. Plant Cell Tiss Org Cult Deng G, Zhang X, Wang T, LinY, Chen X (2008) Chemical composition 15. 97(3): 337-343. and antimicrobial activity of the essential oil of Iris pallid Lam. Chem 30. Industry Forest Prod 28(3): 39-44. Wen B, Wang RL (2010) Pretreatment incubation for culture and cryo preservation of Sabal embryos. Plant Cell Tissue Organ Cult 102: 237- Parang K, Knaus EE, Wiebe LL, SardariS, Daneshtalab M, et al. (1996) 16. 243. Synthesis and antifungal activities of myristic acid analogs. Chem Life 31. Sci 329(11): 475-482. Gonzalez-Arnao MT, William AP, Roca R, Escobar RH, Engelmann F (2008) Development and large scale application of cryopreservation Bonfils JP, Pinguet F, Culine S, Sauvaire Y (2001) Cytotoxicity of iridals, techniques for shoot and somatic embryo cultures of tropical crops. triterpenoids from Iris, on human tumor cell lines A2780 and K562. Plant Cell Tissue Organ Cult 92(1): 1-13. Planta Med 67(1): 79-81.

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How to cite this article: Khalil AL M, Thuraya A z, Nabil AL B. Wild Golden Iris (Iris aurantica) in Syria. Curr Trends Biomedical Eng & Biosci. 2017; 1(5): 00115 555573.